England's four-year, 41-game unbeaten streak is over with Canada upsetting the women's rugby world champions in their WXV Global Series clash in Toronto.
Just over a year after beating Canada to lift the World Cup, the Red Roses were stunned by a late rally from the hosts, who banked a 24-21 win.
England hadn't lost since dropping a 34-31 decision to New Zealand in the 2022 World Cup decider.
The Canadians had already halted England's winning streak a month ago, with the teams playing out a 26-26 draw in Exeter, which was their first match since the 2025 final at Twickenham.
Centre Shoshanah Seumanutafa came off the bench to score the match-winning try in the 77th minute to the delight of 18,621 spectators, which organisers said was a North American record for a stand-alone women's match.
World No.3 Canada looked up for the challenge early as they opened the scoring through fullback Savannah Bauder in the 15th minute.
But quickfire tries to backrower Maddie Feaunati and flyhalf Zoe Harrison just before half-time put England ahead 14-5 at the break.
The points kept flowing as both teams crossed just after play resumed, but DaLeaka Menin's 63rd-minute try kept the door open for Canada.
A raucous home crowd exploded for Seumanutafa's match-winner shortly after, with Canada able to hold on from there and quell England's historic run.
Speaking after the match, England's head coach John Mitchell said he was focused on taking what positives he could out of the loss.
"We're disappointed by the result but there was some very good things in our performance tonight. So we'll stick to our processes," the Kiwi coach said.
"I guess (the loss) frees us up and allows us all to find more.
"I know this group has got more in them but sometimes it requires the context that we've just received to dig deeper and find something more.
"Maybe everyone in the group can probably review their 'why's and re-establish those as well because I'm sure they're changing and evolving as they experience what we're going through."
Mitchell's side will face Canada again next Saturday in Ottawa before completing the away leg of their Global Series fixtures against the USA in New Jersey on October 30.